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Volume 148, Issue 3

Characterization of the replicator region of megaplasmid pTAV3 of and search for plasmid-encoded traits

The GenBank accession number for the sequence reported in this paper is AF390867.

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Abstract

The replicon of the pTAV3 megaplasmid (approx. 400 kb) of has been localized to a 43 kb RI restriction fragment and its entire nucleotide sequence determined. The G+C content of the entire sequence is 66 mol%, which is within the range (62–66 mol%) previously determined for total DNA. ORF1 encodes a replication initiation protein Rep (472 kDa), which shares substantial similarity with putative proteins of the plasmids QpH1 and QpDV, and the replication protein of plasmid pPS10. ORF2, located in the opposite transcriptional orientation to ORF1, encodes a putative protein that shares similarity to a subfamily of ATPases involved in plasmid partitioning. The highest similarity was observed with homologous proteins (RepA) encoded by the family of replicons found in several plasmids of , and spp. The predicted product of ORF3 was similar to AcoR, Nif and NtrC transcriptional activators. A strong incompatibility determinant () was localized between ORF1 () and ORF2 (). The origin of replication of pTAV400 contains a short A+T-rich region and several imperfect palindromic sequences. Curing experiments demonstrated that the megaplasmid bears genes required for growth in minimal media and can therefore be referred to as a mini-chromosome. Megaplasmids pTAV3 of UW1 and pKLW2 of DSM 11073 were found to carry closely related, incompatible replicons. It has been shown that plasmid pORI6 (containing of pTAV3 cloned into plasmid pABW1, which does not replicate in spp.) can be activated not only by pTAV3, but also by pKLW2. Using pORI6, it was demonstrated that replication systems related to pTAV3 are also present in the replicons of JCM 7364, IAM 12816 and DM 12.

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Keyword(s): DR, direct repeat , IR, inverted repeat , Km, kanamycin , MCS, multiple cloning site , mini-replicon , Paracoccus versutus , pTAV3 megaplasmid , Rif, rifampicin and Tc, tetracycline
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2002-03-01
2024-04-19
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Characterization of the replicator region of megaplasmid pTAV3 of Paracoccus versutus and search for plasmid-encoded traitsThe GenBank accession number for the sequence reported in this paper is AF390867.
Microbiology 148, 871 (2002); https://doi.org/10.1099/00221287-148-3-871
/content/journal/micro/10.1099/00221287-148-3-871
/content/journal/micro/10.1099/00221287-148-3-871
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